连翘苷影响TGF-β1/Smad通路抗小鼠肝纤维化作用机制研究  

Forsythin inhibits liver fibrosis by mediating the transforming growth factor-β1/Smad signaling pathway

作　　者：安志强 王清兰 吴中平[1] AN Zhi-qiang;WANG Qing-lan;WU Zhong-ping(College of Basic Medical Science,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《时珍国医国药》2022年第8期1810-1813,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81573810)。

摘　　要：目的探讨连翘苷(Forsythin,FOR)影响TGF-β1/Smad通路抗肝纤维化作用机制。方法选取24只C57BL/6雄性小鼠分成空白对照组、FOR组、模型组、CCL_(4)+FOR组。除空白对照组外,各组小鼠腹腔注射25%的CCL_(4)玉米油造模连续6周;FOR组及CCL_(4)+FOR组腹腔注射给药4周。苏木精-伊红(HE)、天狼星红染色(Sirius Red)、羟脯氨酸(hydroxyproline,Hyp)含量检测、分析血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平评价肝组织和胶原纤维变化;蛋白质印迹法及免疫组化法检测α-平滑肌激动蛋白(α-SMA)、Ⅰ型胶原蛋白(Collagen-Ⅰ);蛋白质印迹法检测TGF-β1、Smad2、Smad 3、p-Smad2、p-Smad 3、Smad7蛋白表达。结果CCL_(4)组纤维沉积加重,ALT,AST显著升高(P<0.01)。CCL_(4)+FOR组小鼠肝细胞变性、坏死及纤维化组织增生程度均较模型组明显减轻,ALT、AST水平、Hyp含量、α-SMA、Collagen-Ⅰ、TGF-β1、p-Smad2,p-Smad3蛋白水平均显著降低,Smad7蛋白表达明显升高(P<0.05,P<0.01)。差异均有统计学意义。结论FOR上调Smad7蛋白表达抑制TGF-β1/Smad通路是其抗纤维化的重要作用机制。Objective To investigate the mechanism of forsythin(FOR)on the effect of TGF-β1/Smad pathway on the liver fibrosis.Methods 24 C57BL/6 male mice were selected and divided into blank control group,FOR group,model group,and CCL_(4)+FOR group.Except for the blank control group,mice in each group were injected intraperitoneally with 25%CCL_(4) corn oil for 6 consecutive weeks;the FOR and CCL_(4)+FOR groups were injected intraperitoneally for 4 weeks.Hematoxylin-Eosin(H&E),Sirius Red staining(Sirius Red),hydroxyproline(Hyp)content detection,analysis of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)level evaluation Changes in liver tissue and collagen fibers;Western blotting and im⁃munohistochemical methods were used to detectα-smooth muscle agonist protein(α-SMA)and type I collagen(Collagen-I);Western blotting was used to detect TGF-β1,Smad2/3,p-Smad2/3,and Smad7 proteins expression.Results The degree of hepatocyte degeneration,necrosis,and fibrotic tissue proliferation in the CCL_(4)+FOR group was significantly reduced compared with the model group.ALT,AST levels,Hyp content,α-SMA,Collagen-I,TGF-β1,p-Smad2/3 levels were significantly reduced,and Smad7 protein expression was significantly increased.The differences are statistically significant.Conclusion FOR up-regulating the expression of Smad7 protein and inhibiting the TGF-β1/Smad pathway is an important mechanism for its anti-fibrosis.

关 键 词：连翘苷 TGF-β1/Smad通路 星状细胞活化 肝纤维化 

分 类 号：R285.5[医药卫生—中药学]